阳虚、阴虚体质唾液miRNA表达谱分析

发布时间：2018-02-20 18:58

本文关键词： 阳虚体质阴虚体质唾液 miRNA 中医体质学　出处：《北京中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:1.探索阳虚、阴虚体质唾液miRNA基因表达谱微观差异,筛选两种偏颇体质微观可分的标志物,探索体质客观研究的新方向。2.分析两种偏颇体质与平和质唾液miRNA基因表达谱微观差异,为下一步通过调体逆转非健康状态体质人群以及诊断和预防疾病提供新的切入点。方法:1.本实验在一般人群中共招募平和体质状态者5人,阳虚体质组未病状态者5例、阴虚体质组未病状态者6例,共计16名不同职业、不同年龄的志愿者。用无菌水漱口以清除口腔内干扰异物,收集待检人员口腔内唾液。分离口腔唾液并提取其中的RNA,进行HumanmiRNAOneArray高通量基因芯片检测。2.通过对miRNA表达谱数据进行分析,筛选出具有差异性表达的miRNA。3.对差异表达miRNA进行靶基因分析,并对靶基因进行功能富集分析。结果:1.本研究发现阳虚体质组与平和体质组相比共有81个差异表达miRNA,包括33个上调差异miRNA和48个下调差异miRNA;阴虚体质组与平和体质组相比共有98个差异表达miRNA,包括35个上调差异miRNA和63个下调差异miRNA;阴虚体质与阳虚体质相比共有167个差异表达miRNA包括51个上调差异miRNA和116个下调差异miRNA。2.通过对差异表达miRNA的靶基因进行KEGG和GO富集分析发现:阳虚与平和对比组存在激素代谢、糖脂代谢、免疫调节、肿瘤等生物学通路的异常表达;阴虚与平和对比组存在炎症反应、免疫调节、肿瘤等生物学通路的异常表达;阳虚与阴虚对比组存在免疫调节、激素代谢、肿瘤等生物学通路的异常表达。结论:1.阳虚与平和、阴虚与平和、阳虚与阴虚组在全基因组层面存在系统性的miRNA表达谱的差异。2.通过靶基因KEGG分析发现阳虚、阴虚体质对比平和质组上调靶基因富集通路中均存在免疫通路的差异表达,显示两种偏颇体质在基因层面可能均存在免疫学通路异常的分子机制。与此同时阳虚对比平和组甲状腺激素、雄性激素等相关激素通路的差异表达,阴虚对比平和组炎性通路、激素相关通路的差异表达,可能是导致阳虚体质虚性低靡特征和阴虚体质虚性亢奋特征的内在原因。3.本研究以人体口腔唾液作为研究对象,选用miRNA表达谱分析作为研究方式,初步构建了阴虚阳虚体质的唾液miRNA表达谱,筛选差异目的基因,分析KEGG、GO富集结果,为探索阳虚、阴虚两种偏颇体质异常表现的分子机制奠定了基础。于此同时将人体口腔唾液作为研究的出发点,也是为了探寻一种更简便可行的方式,为体质微观辨识探寻新方向。
[Abstract]:Objective 1. To explore the microcosmic difference of saliva miRNA gene expression profile between yang-deficiency and yin-deficiency constitution, and to screen two kinds of microcosmic markers of biased constitution. To explore the new direction of objective study of physique. 2. To analyze the microcosmic difference of miRNA gene expression profile between two kinds of biased constitution and placid saliva. This study provides a new entry point for the next step to reverse the unhealthy physique population and to diagnose and prevent diseases. Methods: 1. In the general population, 5 healthy persons were recruited in this experiment, and 5 cases in the Yang deficiency body group were not ill. In Yin-deficiency constitution group, 6 cases were not ill, a total of 16 volunteers of different occupations and ages. Gargle with aseptic water was used to remove foreign body interference in oral cavity. The oral saliva was collected, the oral saliva was isolated and the RNAs were extracted from the saliva, and the HumanmiRNAOneArray high-throughput gene chip was used to detect .2. the data of miRNA expression profile were analyzed. The differentially expressed miRNA.3.The target gene of differentially expressed miRNA was analyzed. The functional enrichment analysis of target gene was carried out. Results 1. This study found that there were 81 differentially expressed miRNAs in Yang deficiency constitution group compared with peaceful constitution group, including 33 up-regulated differential miRNA and 48 down-regulated miRNAs; Yin deficiency constitution group and calmer body group. There were 98 differentially expressed miRNAs, including 35 up-regulated miRNA and 63 down-regulated miRNAs, and 167 differentially expressed miRNA including 51 up-regulated differential miRNA and 116 down-regulated differential miRNA.2. KEGG and go enrichment analysis of target genes expressing differentially expressed miRNA showed that hormone metabolism existed in the control group of Yang deficiency and Ping Ping. Abnormal expression of biological pathways such as glucose and lipid metabolism, immunomodulation, tumor and so on; abnormal expression of inflammatory response, immune regulation, tumor and other biological pathways in Yin-deficiency and calming control groups; immune regulation in Yang deficiency and Yin-deficiency control groups. Conclusion: 1. Yang deficiency and peace, yin deficiency and peace, yang deficiency and yin deficiency have systematic difference of miRNA expression profile on the whole genome level. 2. Yang deficiency is found by KEGG analysis of target gene. There was differential expression of immune pathway in the up-regulated target gene enrichment pathway in calming group compared with that in Yin deficiency group. At the same time, the differential expression of thyroid hormone, androgen and other related hormone pathways in Yang deficiency contrast and Ping Ping group, and the inflammatory pathway in Yin deficiency contrast peace group. The differential expression of hormone related pathways may be the internal cause of the low prevalence of deficiency of Yang and the hyperactivity of deficiency of yin. 3. In this study, oral saliva was used as the research object, and miRNA expression profile analysis was used as the research method. The saliva miRNA expression profile of yin deficiency and yang deficiency constitution was preliminarily constructed, the differentially expressed target genes were screened, and the enrichment results of KEGG go were analyzed in order to explore yang deficiency. The molecular mechanism of two kinds of biased physical abnormalities of yin deficiency has laid a foundation. At the same time taking oral saliva as the starting point is also to explore a more convenient and feasible way to explore a new direction for physique microscopic identification.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R229

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